In some gene editing studies using mice, the delivered plasmid carrying the editing tool (e.g. TALEN) also carries both an HA-tag and GFP. The GFP is separated from the editing tool with a T2A sequence. Why are HA and GFP both needed?
Purifying GFP tagged proteins is labour intensive and usually results in low yields. Adding the HA tag makes it easier and faster to purify these proteins.
To identify the protein, antibody against HA tag can be used for the immunoprecipitation/co-immunoprecipitation and anti-GFP antibody can be used for the detection of the protein level (to avoid cross-linking), or the GFP tagging is used to visualize the localization of the protein and the HA tag is used for immunoprecipitation/purification of the protein due to its lightweight and high specificity.
if the constuct is HAtag/Talen/T2A/GFP : the GFP is used to look in vivo at the transfected cells (%, localisation...), the HA tag is used to detect the editing tool by indirect immunofluorecsnce (IF) or western blot (the level of GFP can be different from the level of expression of the Talen du to difference in protein stability for example)
if the constuct is /Talen/T2A/GFP/HAtag , then the HA tag is not very usefull; GFP can be seen in transfected cells and antibodies against GFP can be used to see the expression but this will give you the level of expression of the GFP not of the talen protein.
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